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A Gierl

Publications and source records attributed to A Gierl.

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Genome organization and transcription in archaebacteria.

The genome organization of the archaebacteria is investigated in three model systems: a) rRNA genes of various archaebacteria, b) a plasmid of 15.6 kb from Sulfolobus acidocaldarius which exists in free or integrated form, c) the 59 kb genome of phage phi H of Halobacterium halobium as a model for the unusual structural variability of DNA in this organism. Several variants of this phage have been isolated, their genomes differ by several insertions, a deletion, and an inversion. The frequent inversion and circularization of a 12 kb segment of DNA appears to be linked to the presence of two copies of an IS element at its flanks. DNA-dependent RNA polymerases have been isolated from a large number of archaebacteria including representatives of 4 families of the novel order Thermoproteales . As shown by immunological methods, they are closely related to those of eukaryotes. Two different types of RNA polymerase exist in the two main branches of the archaebacteria. The role of one component of the enzyme of Thermoplasma acidophilum was elucidated using an in vitro transcription system.

Archaea↗

The role of the components sigma and y of the DNA-dependent RNA polymerase of Lactobacillus curvatus in promotor selection.

An average of 0.44 molecule each of the initiation factor sigma and the RNA polymerase binding protein y and 0.54 molecule of the subunit gamma per molecule of Lactobacillus curvatus DNA-dependent RNA polymerase have been found in the cell. Free factor y displaces sigma from free holo enzyme, E sigma. The formation of a binary complex from Ey, free sigma, and poly[d(A-T)], leads to immediate release of factor y. The release of the sigma factor occurs upon the transition of the binary to a ternary complex. A mixture of E and sigma forms binary complexes with all T7 DNA HpaII restriction fragments. In contrast a mixture of Ey and sigma binds selectively to promoter-containing DNA fragments, indicating that the stimulatory effect of y on transcription is due to an increase in the rate of promoter selection. The same RNA products are synthesised by E sigma and by Ey plus sigma with T7 DNA as template. Thus the nonspecific complexes formed by E sigma and T7 DNA are nonproductive. On the basis of these findings we propose a model for the transcription cycle in Lactobacillus curvatus.

Binding Sites↗

DNA-dependent RNA polymerase of thermoacidophilic archaebacteria.

The component compositions of the DNA-dependent RNA polymerases of the extremely thermophilic, anaerobic sulfur-respiring archaebacteria Thermoproteus tenax and Desulfurococcus mucosus strongly resemble each other but also that of the RNA polymerase of Sulfolobus acidocaldarius suggesting that both organisms belong to the same novel order Thermoproteales, which together with the order represented by Sulfolobus, forms the thermoacidophilic branch of archaebacteria. The component pattern of the RNA polymerase of Thermoplasma acidophilum, which does not belong to this branch, also appears homologous. The archaebacterial type of the DNA-dependent RNA polymerase is thus characterized by 9-10 components yielding a characteristic pattern which resembles that of yeast RNA polymerase A(I). In contrast to the alpha subunit of eubacterial RNA polymerases, the third largest component of archaebacterial RNA polymerases, although similar in size, is present only one per enzyme monomer. The polymerases of T. tenax and D. mucosus, like those previously isolated from other archaebacteria, are completely resistant against 100 microgram/ml rifampicin and streptolydigin. The RNA polymerases of both organisms are highly thermostable. The enzyme from D. mucosus transcribes selectively and almost completely the H strand of phase T7 DNA.

Archaea↗